• 1.上海交通大學(xué)醫(yī)學(xué)院附屬第三人民醫(yī)院普通外科(上海 201900);;
  • 2.安徽蚌埠醫(yī)學(xué)院研究生部(安徽蚌埠 233030);

目的檢測人胃癌KATO-Ⅲ、SGC7901、AGS及MKN-45細(xì)胞株中CD133基因及蛋白的表達(dá)，研究CD133陽性(CD133+)細(xì)胞與CD133陰性(CD133-)細(xì)胞的生物學(xué)特性的差異。方法采用半定量聚合酶鏈反應(yīng)及免疫蛋白印跡法分別檢測KATO-Ⅲ、SGC7901、AGS及MKN-45細(xì)胞株中CD133基因和蛋白的表達(dá); 免疫磁珠細(xì)胞技術(shù)將KATO-Ⅲ細(xì)胞株中CD133+和CD133-細(xì)胞分離純化，對比兩者生長形態(tài)特點、體外增殖能力及分化能力的差異; 采用CCK-8法分析CD133+和CD133-細(xì)胞對不同濃度(0.05、0.10、0.20、0.50、1.00 mg/ml)化療藥物5氟尿嘧啶(5-FU)敏感性的差別。結(jié)果KATO-Ⅲ細(xì)胞中CD133基因及蛋白表達(dá)量均明顯高于SGC7901、AGS及MKN-45細(xì)胞(P lt;0.05)。CD133+細(xì)胞在無血清培養(yǎng)基中呈球形克隆懸浮生長，相比CD133-細(xì)胞具有更強(qiáng)的體外增殖能力及潛在分化能力。5-FU耐藥性實驗中，CD133+細(xì)胞在0.50 mg/ml濃度下的抑制率較CD133-細(xì)胞明顯降低(P lt;0.05)，但其他濃度時二者比較差異無統(tǒng)計學(xué)意義(P gt;0.05)。結(jié)論人胃癌細(xì)胞株KATO-Ⅲ中的CD133+細(xì)胞具有一定增殖、分化及耐藥潛能，CD133可作為胃腫瘤起始細(xì)胞亞群的表面標(biāo)志之一。

引用本文： 陸瑞祺,吳巨鋼,周國才,姜海廣,倪曉春,俞繼衛(wèi),姜波健. 胃癌CD133陽性細(xì)胞的純化及其生物學(xué)特性研究. 中國普外基礎(chǔ)與臨床雜志, 2011, 18(12): 1265-1270. doi: 復(fù)制

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3. Pérez Castillo A, AguilarMorante D, MoralesGarcía JA, et al. Cancer stem cells and brain tumors [J]. Clin Transl Oncol, 2008, 10(5): 262267.
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6. Miraglia S, Godfrey W, Yin AH, et al. A novel fivetransmembrane hematopoietic stem cell antigen: isolation, characterization,and molecular cloning [J]. Blood, 1997, 90(12): 50135021.
7. Kang MK, Kang SK. Tumorigenesis of chemotherapeutic drugresistant cancer stemlike cells in brain glioma [J]. Stem Cells Dev, 2007, 16(5): 837847.
8. O’Brien CA, Pollett A, Gallinger S, et al. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice [J]. Nature, 2007, 445(7123): 106110.
9. Rappa G, Fodstad O, Lorico A. The stem cellassociated antigen CD133 (prominin1) is a molecular therapeutic target for metastatic melanoma [J]. Stem Cells, 2008, 26(12): 30083017.
10. 王曉輝, 李非.胰腺癌干細(xì)胞的研究進(jìn)展 [J]. 中國普外基礎(chǔ)與臨床雜志, 2011, 18(9): 931935.
11. Smith LM, Nesterova A, Ryan MC, et al. CD133/prominin1 is a potential therapeutic target for antibodydrug conjugates in hepatocellular and gastric cancers [J]. Br J Cancer, 2008, 99(1): 100109.
12. Yu JW, Zhang P, Wu JG, et al. Expressions and clinical significances of CD133 protein and CD133 mRNA in primary lesion of gastric adenocacinoma [J]. J Exp Clin Cancer Res, 2010, 29(1): 141151.
13. 吳巨鋼, 陸瑞祺, 張鵬, 等. 胃癌患者外周血CD133 mRNA檢測及其臨床意義 [J]. 中華胃腸外科雜志, 2010, 14(1): 6667.
14. Kelly SE, Di Benedetto A, Greco A, et al. Rapid selection and proliferation of CD133+ cells from cancer cell lines: chemotherapeutic implications [J]. PLoS One, 2010, 5(4): e10035.
15. AlHajj M, Wicha MS, BenitoHernandez A, et al. Prospective identification of tumorigenic breast cancer cells [J]. Proc Natl Acad Sci USA, 2003, 100(7): 39833988.
16. Kume H, Teramoto S, Isurugi K. Invivo dedifferentiation of keratinocytes to epidermal stem cells [J]. Lancet, 2002, 359(9305): 528529.
17. Takenobu H, Shimozato O, Nakamura T, et al. CD133 suppresses neuroblastoma cell differentiation via signal pathway modification [J]. Oncogene, 2011, 30(1): 97105.
18. Wang Z, Li Y, Ahmad A, et al. Pancreatic cancer: understanding and overcoming chemoresistance [J]. Nat Rev Gastroenterol Hepatol, 2011, 8(1): 2733.
19. Chen J, Emara N, Solomides C, et al. Resistance to platinumbased chemotherapy in lung cancer cell lines [J]. Cancer Chemother Pharmacol, 2010, 66(6): 11031111.
20. 張鵬, 俞繼衛(wèi), 姜波健. 腫瘤干細(xì)胞與胃腸道癌發(fā)生和轉(zhuǎn)移 [J]. 中國普外基礎(chǔ)與臨床雜志, 2009, 16(1): 8184.
21. Garvalov BK, Acker T. Cancer stem cells: a new framework for the design of tumor therapies [J]. J Mol Med, 2011, 89(2): 95107.
  1. 1. Bonnet D, Dick JE. Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell [J]. Nat Med, 1997, 3(7): 730737.
  2. 2. Wulf GG, Wang RY, Kuehnle I, et al. A leukemic stem cell with intrinsic drug efflux capacity in acute myeloid leukemia [J]. Blood, 2001, 98(4): 11661173.
  3. 3. Pérez Castillo A, AguilarMorante D, MoralesGarcía JA, et al. Cancer stem cells and brain tumors [J]. Clin Transl Oncol, 2008, 10(5): 262267.
  4. 4. Field M, Alvarez A, Bushnev S, et al. Embryonic stem cell markers distinguishing cancer stem cells from normal human neuronal stem cell populations in malignant glioma patients [J]. Clin Neurosurg, 2010, 57: 151159.
  5. 5. Yu SC, Ping YF, Yi L, et al. Isolation and characterization of cancer stem cells from a human glioblastoma cell line U87 [J]. Cancer Lett, 2008, 265(1): 124134.
  6. 6. Miraglia S, Godfrey W, Yin AH, et al. A novel fivetransmembrane hematopoietic stem cell antigen: isolation, characterization,and molecular cloning [J]. Blood, 1997, 90(12): 50135021.
  7. 7. Kang MK, Kang SK. Tumorigenesis of chemotherapeutic drugresistant cancer stemlike cells in brain glioma [J]. Stem Cells Dev, 2007, 16(5): 837847.
  8. 8. O’Brien CA, Pollett A, Gallinger S, et al. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice [J]. Nature, 2007, 445(7123): 106110.
  9. 9. Rappa G, Fodstad O, Lorico A. The stem cellassociated antigen CD133 (prominin1) is a molecular therapeutic target for metastatic melanoma [J]. Stem Cells, 2008, 26(12): 30083017.
  10. 10. 王曉輝, 李非.胰腺癌干細(xì)胞的研究進(jìn)展 [J]. 中國普外基礎(chǔ)與臨床雜志, 2011, 18(9): 931935.
  11. 11. Smith LM, Nesterova A, Ryan MC, et al. CD133/prominin1 is a potential therapeutic target for antibodydrug conjugates in hepatocellular and gastric cancers [J]. Br J Cancer, 2008, 99(1): 100109.
  12. 12. Yu JW, Zhang P, Wu JG, et al. Expressions and clinical significances of CD133 protein and CD133 mRNA in primary lesion of gastric adenocacinoma [J]. J Exp Clin Cancer Res, 2010, 29(1): 141151.
  13. 13. 吳巨鋼, 陸瑞祺, 張鵬, 等. 胃癌患者外周血CD133 mRNA檢測及其臨床意義 [J]. 中華胃腸外科雜志, 2010, 14(1): 6667.
  14. 14. Kelly SE, Di Benedetto A, Greco A, et al. Rapid selection and proliferation of CD133+ cells from cancer cell lines: chemotherapeutic implications [J]. PLoS One, 2010, 5(4): e10035.
  15. 15. AlHajj M, Wicha MS, BenitoHernandez A, et al. Prospective identification of tumorigenic breast cancer cells [J]. Proc Natl Acad Sci USA, 2003, 100(7): 39833988.
  16. 16. Kume H, Teramoto S, Isurugi K. Invivo dedifferentiation of keratinocytes to epidermal stem cells [J]. Lancet, 2002, 359(9305): 528529.
  17. 17. Takenobu H, Shimozato O, Nakamura T, et al. CD133 suppresses neuroblastoma cell differentiation via signal pathway modification [J]. Oncogene, 2011, 30(1): 97105.
  18. 18. Wang Z, Li Y, Ahmad A, et al. Pancreatic cancer: understanding and overcoming chemoresistance [J]. Nat Rev Gastroenterol Hepatol, 2011, 8(1): 2733.
  19. 19. Chen J, Emara N, Solomides C, et al. Resistance to platinumbased chemotherapy in lung cancer cell lines [J]. Cancer Chemother Pharmacol, 2010, 66(6): 11031111.
  20. 20. 張鵬, 俞繼衛(wèi), 姜波健. 腫瘤干細(xì)胞與胃腸道癌發(fā)生和轉(zhuǎn)移 [J]. 中國普外基礎(chǔ)與臨床雜志, 2009, 16(1): 8184.
  21. 21. Garvalov BK, Acker T. Cancer stem cells: a new framework for the design of tumor therapies [J]. J Mol Med, 2011, 89(2): 95107.